The major goals of our research have been to identify the mechanisms of activation of the farnesoid X receptor (FXR) and to define its role in regulating metabolic pathways. To this end, we have identified four human and murine FXR transcripts derived from a single gene that encode four different protein isoforms. Our identification of a four amino acid motif (MYTG), located immediately adjacent to the DNA binding domain of two of the four isoforms, dramatically altered our thinking about this transcription factor; new and exciting data suggest that the presence of this motif affects transactivation of certain hepatic and adrenal genes. We have recently discovered that activated FXR has a pronounced effect on glucose metabolism in mice and is highly protective against the acute hepatoxicity produced by toxic xenobiotics and from the toxic effects of IPS (lipopolysaccharide). We have also recently obtained evidence that FXR regulates a number of steroidogenic genes, suggesting a functional role for FXR in the adrenal cortex. Based on these data, generated during the current grant period, we propose to conduct mechanistic studies to elucidate the roles of FXR in i) the control of plasma glucose levels and hepatic glucose metabolism, ii) the control of plasma and hepatic lipid levels, iii) protection of the liver from damage induced by xenobiotics such as acetaminophen, iv) protecting mice from endotoxin shock and bacterial infection and v) the regulation of target genes in adrenal steroidogenic cells. These studies will be aided by the availability of FXR transgenic and FXR-/- mice, and mice lacking FXR in the liver or intestine, and adenovirus expressing individual FXR isoforms. To complement these approaches, mechanistic studies will be conducted to elucidate the function of the MYTG motif present in two of the four FXR isoforms. Taken together, these studies will identify novel regulatory mechanisms by which FXR isoforms activate transcription. In addition, these studies will elucidate the role of FXR in glucose, steroid and drug metabolism and in resistance to endotoxin.